Project Summary/Abstract The direct analysis of "raw" genomic DNA offers an unfiltered and relatively unbiased view of any genome obviating need for clone libraries and PCR amplicons. This advantage is greatly potentiated by the analysis of large numbers of single DNA molecules for creation of voluminous data sets. As such, the development of a scheme for acquisition of sequence information is proposed using large genomic DNA molecules that will be optically barcoded and analyzed for sequence content, offering resolution approaching that of resequencing. Because large DNA molecules are analyzed, this sequence acquisition scheme obtains information from heterochromatic regions, pinpoints structural variants in human genomes, and characterizes aberrations associated with cancer genomes. This scheme also offers opportunities for linking sequence data from emerging sequencing platforms.